中国实验动物学报Issue(4)：359-364,6.DOI:10.3969/j.issn.1005-4847.2015.04.006
Rab13通过影响 PKA 活性调节 occludin 及 F-actin在大鼠血睾屏障的分布
Rab13 regulates the distribution of occludin and F-actin at the blood-testis barrier via PKA activity in rats
摘要
Abstract
Objective To investigate the regulation of blood-testis barrier by Rab13-PKA pathway in rats.Meth-od First, shRNA vector targeting at Rab13 was constructed and then the Rab13 shRNA was transfected into the rat testis by injection.Western blot was used to detect the knock-down effect of Rab13 and the expression of blood-testis barrier ( BTB) constituent proteins.PKA activity was detected by autoradiography and scintillation counting.Further, immunoflu-orescence analysis and phalloidin staining were applied to observe the distribution of occludin and F-actin, respectively. Results The expression level of Rab13 in the testis was reduced by approximately 70%after transfection of Rab13 shRNA as compared with the non-targeted control group ( P<0.01 ) , while the expression of BTB constituent proteins remained unchanged.PKA activity was significantly increased after Rab13 RNAi transfection (P<0.01).The distribution of occlu-din at BTB was remarkably increased after Rab13 RNAi silencing around stage VIII but not at other stages of the seminifer-ous epithelial cycle.The assembly of F-actin at BTB was also intensified in Rab13-silenced testis.Both the changes of dis-tribution of occludin and F-actin induced by Rab13 shRNA were found to be antagonized by the PKA specific inhibitor H89.Conclusions Rab13 can modulate the distribution of occludin and F-actin at the blood-testis barrier in rats by regu-lating PKA activity, which may participate in the regulation of BTB function.关键词
Rab13/血睾屏障/蛋白激酶AKey words
Rab13/Blood-testis barrier/Protein kinase A/Rats分类
生物科学引用本文复制引用
宿文辉,贾晓宇,孟晓娜..Rab13通过影响 PKA 活性调节 occludin 及 F-actin在大鼠血睾屏障的分布[J].中国实验动物学报,2015,(4):359-364,6.基金项目
国家自然科学基金项目（批准号81100462，81370756）。 （）
